To analyze the mechanisms of HCV drug resistance by using chimeric HCV cell culture system

• Project/Area Number: 25860513
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Gastroenterology
• Research Institution: Hokkaido University
• Principal Investigator: SUDA Goki 北海道大学, 医学(系)研究科(研究院), 特任助教 (20447460)
• Co-Investigator(Renkei-kenkyūsha): SAKAMOTO Naoya 北海道大学, 大学院医学研究科, 教授 (10334418)
• Project Period (FY): 2013-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: ＨＣＶ / ＩＦＮ / 薬剤耐性ウイルス / HCV

Outline of Final Research Achievements

The mechanism of HCV resistance to IFN has not been wellunderstood. We conducted homologous recombination between HCV-2b and JFH1 and ensured that the chimeric virus could be long-cultured. Then we long-cultured HCV-2b/JFH1 chimeric virus (C3) with or without IFN and compared virus kinetics between the two groups, and tried to extract IFN resistant clone. Supernatant HCV of C3 decreased immediately. However, 6 weeks after IFN treatment, replication of one C3 clone increased . Comparison of amino acid sequences revealed two sequence differences in structure region. Next we constructed these two sequence differences substituted C3 clone (IFNrC3) and compared IFN sensitivity between IFNrC3 and C3 by transfection into cells and subsequently IFN treatment. The newly constructed IFNrC3 showed resistance to IFN.
Additionally, we investigated prevalence of resistant HCV against DAA in japan by using NGS. The existence of resistant virus did not affect treatment outcome in TPV based therapy.

Research Products

• [Journal Article] EGFR inhibitors eliminate cancer stem-like cells in esophageal squamous cell carcinoma by suppressing epithelial-mesenchymal transition (2015)
  • Author(s): １．Fumiyuki Sato, Yoshimasa Kubota, Mitsuteru Natsuizaka, Osamu Maehara, Yutaka Hatanaka, Katsuji Marukawa, Katsumi Terashita, Goki Suda, Shunsuke Ohnishi, Yuichi Shimizu, Yoshito Komatsu, Shinya Ohashi, Shingo Kagawa, Hideaki Kinugasa, Kelly Whelan, Anil Rustgi, Hiroshi Nakagawa, and Naoya Sakamoto
  • Journal Title: Cancer Biology & Therapy Volume: Epub ahead of print
  • Peer Reviewed
• [Journal Article] Human amnion-derived mesenchymeal stem cell transplantation ameliorates dextran sulfate sodium-induced severe colitis in rats. (2015)
  • Author(s): ２．Onishi R, Ohnishi S, Higashi R, Watari M, Yamahara K, Okubo N, Nakagawa K, Katsurada T, Suda G, Natsuizaka M, Takeda H, Sakamoto N
  • Journal Title: Cell Transplantation Volume: Epub ahead of print
  • NAID: 120005690898
  • Peer Reviewed
• [Journal Article] Intratumoral artery on contrast-enhanced computed tomography imaging: differentiating intrahepatic cholangiocarcinoma from poorly differentiated hepatocellular carcinoma (2015)
  • Author(s): ３．Tsunematsu S1, Chuma M, Kamiyama T, Miyamoto N, Yabusaki S, Hatanaka K, Mitsuhashi T, Kamachi H, Yokoo H, Kakisaka T, Tsuruga Y, Orimo T, Wakayama K, Ito J, Sato F, Terashita K, Nakai M, Tsukuda Y, Sho T, Suda G, Morikawa K, Natsuizaka M, Nakanishi M, Ogawa K, Taketomi A, Matsuno Y, Sakamoto N
  • Journal Title: Abdom Imaging. Volume: Epub ahead of print
  • Peer Reviewed
• [Journal Article] Serum granulysin levels as a predictor of serious telaprevir-induced dermatological reactions. (2015)
  • Author(s): Suda G, Yamamoto Y, Nagasaka A, Furuya K, Kudo M, Chuganji Y, Tsukuda Y, Tsunematsu S, Sato F, Terasita K, Nakai M, Horimoto H, Sho T, Natsuizaka M, Ogawa K, Ohnishi S, Chuma M, Fujita Y, Abe R, Taniguchi M, Nakagawa M, Asahina Y, Sakamoto N
  • Journal Title: Hepatol Res Volume: 印刷中 Issue: 8 Pages: 837-45
  • DOI: 10.1111/hepr.12421
  • NAID: 120005820706
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Heat shock factor 1 accelerates hepatocellular carcinoma development by activating nuclear factor κB/mitogen-activated protein kinase. (2014)
  • Author(s): Chuma M, Sakamoto N, Nakai A, Hige S, Nakanishi M, Natsuizaka M, Suda G, Sho T, Hatakaka K, Matsuno Y, Yokoo H, Kamiyama T, Taketomi A, Fujii G, Tashiro K, Hikiba Y, Fujimoto M, Asaka M, Maeda S
  • Journal Title: Carcinogenesis Volume: Epub ahead of print Issue: 2 Pages: 272-281
  • DOI: 10.1093/carcin/bgt343
  • Peer Reviewed
• [Journal Article] Hepatosplenic Gamma-delta T-cell Lymphoma Associated with Epstein-Barr Virus (2014)
  • Author(s): ６．Seiji Tsunematsu, Mitsuteru Natsuizaka, Hiromi Fujita, Noriyuki Otsuka, Katsumi Terashita, Fumiyuki Sato, Tomoe Kobayashi, Masato Nakai, Yoko Tsukuda, Hiromasa Horimoto, Takuya Sho, Goki Suda, Mitsuru Nakanishi, Satoshi Hashino, Makoto Chuma, Naoya Sakamoto
  • Journal Title: Intern Med Volume: 53(18) Pages: 2079-2082
  • NAID: 130004687645
  • Peer Reviewed
• [Journal Article] Telaprevir:副作用とその対策 (2014)
  • Author(s): 須田剛生、坂本直哉
  • Journal Title: Internal Medicine Volume: ６７ Pages: 855-860
• [Journal Article] インターフェロン少量長期療法の実際と新薬時代における役割 (2014)
  • Author(s): 須田剛生、坂本直哉
  • Journal Title: 内科 Volume: 113 Pages: 681-685
• [Journal Article] C型肝炎ウイルス蛋白によるシグナル伝達制御 (2013)
  • Author(s): 須田剛生、坂本直哉
  • Journal Title: 分子消化器病 Volume: 10 Pages: 68-72
• [Journal Article] IL6を介したC型慢性肝炎のインターフェロン治療抵抗性機構 (2013)
  • Author(s): 須田剛生、坂本直哉
  • Journal Title: 消化器内科 Volume: 56 Pages: 421-426
• [Presentation] DAA併用療法におけるDAA耐性ウイルス/治療予測因子の検討 (2014)
  • Author(s): 須田剛生 古家乾 坂本直哉
  • Organizer: 肝臓病学会東部会
  • Place of Presentation: 京王プラザホテル （東京 新宿）
  • Year and Date: 2014-11-29
• [Presentation] Ｃ型肝炎は完治できる時代へ (2014)
  • Author(s): 須田剛生
  • Organizer: 日本臨床検査医学学術集会
  • Place of Presentation: 福岡国際会議場 （福岡県 博多）
  • Year and Date: 2014-11-24
• [Presentation] Serum granulysin levels as a predictor of serious telaprevir-induced dermatological reaction (2014)
  • Author(s): Goki Suda1 ,Yoko Tsukuda1, Seiji Tsunematsu1, Fumiyuki Satou1, Katsumi Terasita1, Takuya Sho 1, Masato Nakai1, Mitsuteru Natsuizka1, Makoto Chuma 1, Naoya Sakamoto
  • Organizer: 65th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: Boston (USA)
  • Year and Date: 2014-11-09
• [Presentation] Analysis of First and Second Generation Protease Inhibitor Combination Therapy in Patients with Chronic Hepatitis C Virus Infection (2014)
  • Author(s): 須田剛生 永坂敦 坂本直哉
  • Organizer: 日本消化器病関連学会週間
  • Place of Presentation: 神戸国際展示場 （兵庫県 神戸）
  • Year and Date: 2014-10-23
• [Presentation] C型慢性肝炎に対するDAA療法の治療予測因子・薬剤耐性ウイルスの検討 (2014)
  • Author(s): 須田剛生 中馬 誠 坂本直哉
  • Organizer: 肝臓病学会総会
  • Place of Presentation: ホテルニューオータニ （東京都 千代田区)
  • Year and Date: 2014-05-29
• [Presentation] プロテアーゼ阻害剤併用療法の治療予測因子・薬剤耐性ウイルスの検討 (2014)
  • Author(s): 須田剛生 中馬 誠 坂本直哉
  • Organizer: 消化器病学会総会
  • Place of Presentation: 東京国際フォーラム (東京都 千代田区）
  • Year and Date: 2014-04-24
• [Presentation] Serum granulysin level is correlated with severity of telaprevir induced dermatological adverse reactions and could be a useful predictor (2014)
  • Author(s): Goki Suda , Yoshiya Yamamoto, Astusi Nagasaka, Ken Furuya, Yoko Tsukuda, Seiji Tsunematsu, Fumiyuki Satou, Katsumi Terasita, Keita Horimoto, Takuya Sh Masato Nakai, Mitsuteru Natsuizka, Makoto Chuma , Naoya Sakamoto
  • Organizer: Asian Pacific Association for The Study of the Liver
  • Place of Presentation: Brisbane Convention & Exhibition Centre（Brisbene）, Australia
• [Presentation] Establishment of Interferon resistant genotype 2b/JFH-1 chimeric Hepatitis C virus cell culture system (2013)
  • Author(s): Goki Suda, Yoko Tsukuda, Mitsuteru Natsuizaka, Makoto Chuma, Naoya Sakamoto
  • Organizer: 64th. Annual Meeting of American Association for the Study of Liver Diseases
  • Place of Presentation: Walter E. Washington Convention Center （Wachington DC）, USA
• [Presentation] Telaprevir/Peg-IFN/RBV3剤併用療法の治療効果および副作用予測因子の解析 (2013)
  • Author(s): 須田剛生 中馬誠 坂本直哉
  • Organizer: 日本消化器病関連学会週間
  • Place of Presentation: グランドプリンス高輪ホテル（東京）